High-throughput and Sensitive Immunopeptidomics Platform Reveals Profound Interferonγ-Mediated Remodeling of the Human Leukocyte Antigen (HLA) Ligandome
| Autor: | Michal Bassani-Sternberg, Julien Racle, Chloe Chong, Fabio Marino, Markus Müller, Roy Thomas Daniel, HuiSong Pak, George Coukos, David Gfeller |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Proteomics
0301 basic medicine T-Lymphocytes Human leukocyte antigen Ligands Biochemistry Cell Line Analytical Chemistry Interferon-gamma 03 medical and health sciences 0302 clinical medicine Neoplasms medicine Humans Interferon gamma Molecular Biology B-Lymphocytes/metabolism Histocompatibility Antigens Class I/metabolism Histocompatibility Antigens Class II/metabolism Interferon-gamma/pharmacology Neoplasms/metabolism Peptides/metabolism Proteomics/methods T-Lymphocytes/metabolism B-Lymphocytes Antigen processing Chemistry Histocompatibility Antigens Class I Histocompatibility Antigens Class II Technological Innovation and Resources Cancer medicine.disease Cell biology 030104 developmental biology Cell culture 030220 oncology & carcinogenesis Ovarian cancer cells Peptides medicine.drug |
| Zdroj: | Molecular & cellular proteomics, vol. 17, no. 3, pp. 533-548 Molecular & Cellular Proteomics : MCP |
| Popis: | Comprehensive knowledge of the human leukocyte antigen (HLA) class-I and class-II peptides presented to T-cells is crucial for designing innovative therapeutics against cancer and other diseases. However methodologies for their purification for mass-spectrometry analysis have been a major limitation. We designed a novel high-throughput, reproducible and sensitive method for sequential immuno-affinity purification of HLA-I and -II peptides from up to 96 samples in a plate format, suitable for both cell lines and tissues. Our methodology drastically reduces sample-handling and can be completed within five hours. We challenged our methodology by extracting HLA peptides from multiple replicates of tissues (n = 7) and cell lines (n = 21, 10 8 cells per replicate), which resulted in unprecedented depth, sensitivity and high reproducibility (Pearson correlations up to 0.98 and 0.97 for HLA-I and HLA-II). Because of the method's achieved sensitivity, even single measurements of peptides purified from 10 7 B-cells resulted in the identification of more than 1700 HLA-I and 2200 HLA-II peptides. We demonstrate the feasibility of performing drug-screening by using ovarian cancer cells treated with interferon gamma (IFNγ). Our analysis revealed an augmented presentation of chymotryptic-like and longer ligands associated with IFNγ induced changes of the antigen processing and presentation machinery. This straightforward method is applicable for basic and clinical applications. |
| Databáze: | OpenAIRE |
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